metastasis lecture and seminar

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Progression des tumeurs et Interactions
tumeur-hôte
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Propriétés essentielles des cellules cancéreuses
Autosuffisance en signaux de croissance
Resistance à lapoptose
Résistance aux signaux anti-croissance
Angiogénèse
Invasion et métastases
Potentiel de division illimité
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Les tumeurs sont

• Monoclonales
• mais néanmoins
• hétérogènes

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Monoclonalité des tumeurs
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Hétérogénité des cellules tumorales
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Mécanismes de progression

• hétérogénéité génétique des cellules tumorales à
  cause de linstabilité
• de lADN (défaillance des mécanismes réparateurs)
• chromosomique
• sélection de nouveaux sous-clônes avec une plus
  grande capacité de croissance (locale ou
  métastatique)

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Progression des tumeurs
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Cancer stem cells
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Cancer stem cells (CSC) distinct from the cell
of origin CSC do not necessarily originate from
the transformation of normal stem cells CSC may
arise from restricted progenitors or more
differentiated cells that have acquire
self-renewal properties. Parallels between
self-renewal programs of CSC and normal stem
cells?
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Glioblastome multiforme
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Glioblastome multiforme (notez laspect infiltrati
f avec une forte vascularisation, des cellules
hautement pléomorphes et des zones de necrose)
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Isolation and propagation of brain tumor stem
cells in culture
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Assays for tumor cells
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Definition of brain tumor stem cells Cancer
initiating ability upon orthotopic implnatation
(tumors should be a phenocopy of the tumor of
origin) Extensive self-renewal ability,
demonstrated either ex-vivo (by showing both
sequencial-clonogenic and population-kinetic
analyses) or in vivo (by serial, orthotopic
transplantation) Karyotypic or genetic
alterations Aberrant differentation
properties Capacity to generate non-tumorigenic
end cells Multilineage differentiation capacity
(facultative)
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Interactions tumeur-hôte
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Endothelial cell
Tumor cell
leukocyte
fibroblast
Tumeur composition as a function of growth
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Carcinome cellules tumorales et cellules
stromales
Stroma
Tumeur
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Carcinome canalire invasif du sein
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Carcinome canalaire invasif du sein
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Carcinome lobulaire invasif du sein
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Interactions tumeur-hôte
Le cancer est composé de cellules cancereuses et
de cellules normales provenant du stroma
avoisinant, comprenant les cellules vasculaires
endotheliales, les(myo)fibroblastes et les
cellules inflammatoires - on peut considérer
le cancer comme un organe dans lorgane
hôte - le cancer peut aussi être considéré
comme une plaie qui ne guérit jamais
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vaisseau sanguin
kinines
complément
coagulation
fibrinolyse
anticorps
endothelium
chimiokines
macrophage
mastocyte
C5a
voie alterne
voie classique
complement
complexes immuns
microbes
anticorps circulants
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Plaie
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Réaction du stroma et réponse inflammatoire
(recrutement de leukocytes, production de
facteurs de croissance, remaniement de la
matrice extracellulaire, angiogénèse,
proliferation cellulaire, etc).
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Réparation du tissu atténuation de
linflammation
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Réparation du tissu atténuation de
linflammation
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Réparation du tissu atténuation de
linflammation
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Résolution - cicatrice
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Intégrité du tissu
Epithélium normal
Stroma normal
Précancer
CANCER
STROMA TUMORAL
INVASION
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Progression des tumeurs

• La capacité denvahir les tissus est considérée
  comme un signe de progression tumorale. Lorsque
  les cellules tumorales pénètrent la membrane
  basale, elles induisent une réaction du stroma du
  tissu hôte cette réaction stromale peut refléter
  une tentative de défense mais va en général
  servir les besoins des cellules tumorales.

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Progression des tumeurs

• les récidives
• sont mois bien différenciés
• poussent plus rapidement
• répondent moins au traitement
• métastatisent plus facilement
• sont plus agressives

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Interactions tumeur-hôte
Angiogenèse tumorale Mécanismes identiques à
ceux qui régulent langiogenèse physiologique Le
principal facteur stimulant de langiogenèse
dans les tumeurs est lhypoxie tissulaire
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Angiogenèse tumorale Angiogenic switch
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L angiogénèse
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Hypoxie
ANG1 ANG2 VEGF A
Tie2
Tie2
VEGF-R1/R2
PC
Recrutement and interaction with peri-endothelial
cells (pericytes and smooth muscle cells)
Loosening of contacts with the ECM and
support cells
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Angiogenèse
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Lymphangiogenèse
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VEGF-C
Sema3
Lymphatic endothelial cells
Integrins
NRP-2
Plexin
VEGF-R2 VEGF-R3
Regulation of endothelial cell growth
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CCL21
VEGF-C
Vaisseau lymphatique
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Croissance invasive
Interactions tumeur-hôte

• détachement intercellulaire
• attachement à la matrice extracellulaire
• dégradation de la matrice extracellulaire
• migration des cellules tumorales

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Endothelial cell
Fibroblast
GF
Lymphocyte
ECM
Epithelial cells
Carcinoma cells
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Endothelial cell
fibroblast
lymphocyte
Growth factors cytokines
Contact-de- pendent signal exchange
Cancer cells
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Endothelial cell
fibroblast
lymphocyte
Tissue remodeling
Stimulation of migration and growth
Tumor cells
Migration invasion survival proliferation
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Facteurs de croissance/cytokines sequesterés
dans la MEC
TGF-b contrôle de la croissance cellulaire,
induction de linvasivité, promotion de
langiogenèse transition épitheliale- mesenchyma
teuse desmoplasie fibroplasie) bFGF --
angiogenèse IGF-1 -- survie, prolifération VEGF
angiogenèse Angiopoietin-1 angiogenèse PDGF
proliferation, desmoplasie HGF/SF
proliferation, migration, invasion
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Interactions tumeur-hôte
Cellule tumorale
fibroblaste
différentiation
myofibroblaste
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Interactions tumeur-hôte
Cellule tumorale invasive
Cellule tumorale potentielement invasive
PDGF IFN-g
Signaux pro-invasion
Complexe TGF-b latent
macrophage
MMP et autres protéases dérivées du tissu hôte
TGF-b actif
fibroblaste myofibroblaste
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Dualité du Rôle de TGF-b
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Voies de signalisation de TGF-b
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Altération de léquilibre de signalisation vers
la tumorigénèse
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Modulation de la voie de signalisation SMAD dans
les tumeurs
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Stamenkovic_Figure3_
Angiogenesis
Recruitment, proliferation, activation
VEGF-A
Hematopoietic and endothelial precursor cell
(HPC, EPC)
Lymphangiogenesis
VEGFR1
VEGFR3
Tumor cell
VEGFC, VEGFD
Blood vessel
Lymphatic vessel
Proliferation/differentiation
Normal epithelia
TGF-b
BM
EMT/Invasion
Fibroblast
ECM remodeling (MMP9, uPA, cathepsins)
Activated fibroblast
Collagen fibres
Myofibroblast (CAF)
Release of sequestred growth factors (IGF1,
TGF-b, PDGF, VEGF, bFGF, HGF/SF)
Granulocyte
Monocyte/TAM
EPC recruitment
SDF-1
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Fibroblast
Lymphocyte
Endothelial cell
proTGF-b
FGFR
ECM
bFGF
PG
VEGF
uPA
MMP/TIMP
SF/HGF
RGD/Integrins
PG
VEGFR
uPAR
TGFbR
C-Met
cadherins
PG
MTiMMP
FAK
HB-EGF
PI(3)K
Carcinoma cell
EGFR
catenin
MLCK
ERK
Akt
Rho
Rac
Cdc42
Cytoskeletal and ECM remodelling Actin
polymerization and myosin contraction Invadopodia
protrusion Attachment/detachment
Gene transcription Bypass/override of cell cycle
checkpoints
Suppression of apoptosis and avoidance of anoikis
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chimiothérapie
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chimiothérapie
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radiothérapie
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chimiothérapie
thérapie ciblée
thérapie ciblée
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chimiothérapie
thérapie ciblée
thérapie ciblée
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chimiothérapie
thérapie ciblée
thérapie ciblée
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Molecular mechanisms of Cancer metastasis
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Propriétés essentielles des cellules cancéreuses
Autosuffisance en signaux de croissance
Resistance à lapoptose
Résistance aux signaux anti-croissance
Angiogénèse
Invasion et métastases
Potentiel de division illimité
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Transformation
Cellule normale Cellule en voie de
transformation Cellule cancéreuse
Réseau de signalisation Déréglement du
réseau Nouveau réseau
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Progression des tumeurs
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Tumor development and progression
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Cancer metastasis pathways
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Lung cancer metastasis
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Liver metastases
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Une micrométastase ganglionnaire
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Lung metastasis of colon carcinoma
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Tumor Metastasis
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Mutation in relevant genes Clonal expansion and
genetic instability Inhibition of
apoptosis Angiogenesis and selection of variants
Primary tumor
Detachment of cells from primary mass Inhibition
of anoikis Alteration of contacts between tumor
and ECM Degradation of ECM and migration
Invasion
Degradation of vascular basement
membrane Migration through endothelium
Intravasation
Evasion of immune surveillance Resistance to
mechanical stress Adhesion to platelets Adhesion
to endothelium of distant organs
Circulate in blood and lymph
Proliferation on endothelial cells Disruption of
endothelial basement membrane Endothelial cell
apoptosis
Extravasation
Metastatic tumor
Establishment of a relationship with host
tissue stroma conducive to tumor colony formation
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Tumor-host interactions
Proteolysis
Adhesion
Resistance to apoptosis
Sustained angiogenesis
migration
metastasis
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STEP 1 Detachment
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Changes of adhesion properties in transformed
cells
Growth promotion migration
Growth control survival
cadherin
Inactive Proteoglycan (e.g.CD44)
Selectin ligands
a5b1 integrin
avb3 integrin
Active proteoglycan
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Changes of proteolytic properties in transformed
cells
Growth promotion migration
Growth control survival
Cell surface and soluble proteases
Factors that stimulate protease secretion by
stroma
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Stamenkovic_Figure2_
Selectin ligands
CD44 inactive
CD44v3
cMet
N-cadherin
GAG chains
proHB-EGF
E-cadherin
FGFR
MMP7
ErbB4
cMet
a5
b1
a3
b1
BM
av
Integrins
b 3
b4
a6
Integrins
MMP14
MMP15
ErbB2
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STEP 2 Migration/Invasion of basement membrane
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Endothelial cell
Fibroblast
GF
Lymphocyte
ECM
Epithelial cells
Carcinoma cells
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Regulation of the actin cytoskeleton by Rho
GTPases
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Regulation of Rho GTPases
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Rho-GTP
GEF
GAP
Rho-GDP
Rho-GTP
Rho-GDP
Rho-GDP
Rho-GTP
Inactive GEF
Inactive GEF
Rho-GTP
Rho-GDP
P
PTP
P
PTP
Active GEF
Active GEF
P
P
Rho-GDP
Rho-GTP
Rho-GTP
Rho-GDP
Model for thepotential regulation of Rho
activity by tyrosine phosphatases
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Endothelial cell
fibroblast
lymphocyte
Growth factors cytokines
Contact-de- pendent signal excahnge
Cancer cells
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Endothelial cell
fibroblast
lymphocyte
Tissue remodeling
Stimulation of migration and growth
Tumor cells
Migration invasion survival proliferation
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STEP 3 Intravasation Proteolysis of basement
membrane (MMPs and other enzymes) Disruption of
endothelium Penetration into the circulation
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Stamenkovic_Figure1_
Invasive carcinoma
Detachment
In situ carcinoma
Normal epithelia
BM
Invasion
BM degradation
Migration
No proliferation
Dormant metastases
Intravasation
Circulation
Extravasation
Proliferation, angiogenesis, microenvironment
activation
Growing metastases
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lymphocyte
fibroblast
Basement membrane
endothelium
Growth factors
ECM proteins -collagen
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lymphocyte
fibroblast
leukocyte
Basement membrane
endothelium
Growth factors
ECM proteins -collagen
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STEP 4 Survival within the circulation -resistanc
e to shear stress -adhesion to platelets (some
tumor types) -sialylation (masking) of
immunogenic epitopes -proteolytic cleavage of
adhesion receptors implicated in interactions
with cytotoxic lymphocytes -proteolytic cleavage
of death receptors (Fas, TNFR)
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P-selectin
platelet
Sulfated,sialylated lactosaminoglycans
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lymphocyte
fibroblast
Basement membrane
endothelium
Growth factors
ECM proteins -collagen
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STEP 5 Immobilization and proliferation
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lymphocyte
fibroblast
Basement membrane
endothelium
Growth factors
ECM proteins -collagen
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STEP 6 Extravasation
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lymphocyte
fibroblast
Growth factors
ECM proteins -collagen
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STEP 7 Establishment of a relationship with the
host tissue stroma that is favorable to tumor
growth
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Endothelial cell
fibroblast
lymphocyte
Growth factors cytokines
Contact-de- pendent signal excahnge
Cancer cells
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Endothelial cell
fibroblast
lymphocyte
Tissue remodeling
Stimulation of migration and growth
Tumor cells
Migration invasion survival proliferation
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Growth factors/cytokines sequestered in the ECM
TGF-b growth control, induction of
invasiveness, EMT, angiogenesis bFGF --
angiogenesis IGF-1 -- proliferation,
survival VEGF angiogenesis Angiopoietin-1
angiogenesis PDGF proliferation,
desmoplasia HGF/SF proliferation, migration,
invasion
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Fibroblast
Lymphocyte
Endothelial cell
proTGF-b
FGFR
ECM
bFGF
PG
VEGF
uPA
MMP/TIMP
SF/HGF
RGD/Integrins
PG
VEGFR
uPAR
TGFbR
C-Met
cadherins
PG
MTiMMP
FAK
HB-EGF
PI(3)K
Carcinoma cell
EGFR
catenin
MLCK
ERK
Akt
Rho
Rac
Cdc42
Cytoskeletal and ECM remodelling Actin
polymerization and myosin contraction Invadopodia
protrusion Attachment/detachment
Gene transcription Bypass/override of cell cycle
checkpoints
Suppression of apoptosis and avoidance of anoikis
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Problems facing the metastasizing tumor cell
Event Underlying
mechanism
Adhere to endothelium Adhesion Penetrate
the endothelial barrier, i.e.basement
membrane Proteolysis Overcome the barrier
presented by ECM proteins Proteolysis Use the
ECM as a scaffold to migrate on Adhesion Gain
access to ECM-sequestered growth
factors Proteolysis SURVIVE and
PROLIFERATE Adhesion, proteolysis,
angiogenesis and intrinsic resistance to
apoptosis
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Adhesion receptors relevant to cancer progression
Adhesion receptor class Normal function Cancer
cells Integrins adhesion to ECM change
of (gt20 a and 8 b chains) growth
control repertoire survival signals growth
signals migration Cadherins cell-cell
interaction expression cell
junctions downregu- growth
control lation Proteoglycans (e.g.CD44) adhesion
to ECM migration growth factor re- MMP
anchors gulation growth signals Selectins/s
electin ligands leukocyte rolling adhesion
to on endothelium endothelium and
platelets
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Proteolytic enzymes relevant to cancer progression
Enzyme class Function Matrix
metalloproteinases (MMPs) ECM protein
degradation Latent growth factor
activation Cell surface receptor
regulation A disintegrin and metalloproteinase (A
DAMs) Latent growth factor activation Cell
surface receptor cleavage Adhesion uPA/uPAR
/plasmin regulation of coagulation MMP
activation Latent growth factor
cleavage Cathepsins Expression increases
with tumor progression
tumor stromal interface
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MMP structure
Minimal domain MMPs (MMP7, matrilysin-2) Simpl
e hemopexin-domain MMPs Gelatin-binding MMPs
(MMP-2, -9) Transmembrane MMPs (MMP14, 15,
16)
Pre Pro Catalytic Zn
Hemopexin
Pre Pro Catalytic Zn
S S
Fn repeats
Hemopexin
Pre Pro Catalytic
Zn
S S
Hemopexin Tm Cyt
Pre Pro Catalytic Zn
S S
Hemopexin
Control of MMP activity by TIMPs
S S
TIMP
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Proteolysis in Cancer Progression
ECM degradation Growth Factor Cell surface
receptor release and activation cleavage
Creation of a path Generation of
breakdown products with significant biological
activity
Activation of migration, invasion, growth and
survival programs
Regulation of receptor function
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MMP substrates
ECM proteins Collagens MMP-1, 2, 7, 9,
13 Gelatin MMP-2, 9 Laminin,
fibronectin MMP-1, 2, 3, 7, 9 Entactin MMP-3,
7, 11, 13 Perlecan MMP-7 Tenascin MMP-2,
3, 7, 9 Growth factors Latent TGF-beta MMP-2,
-9 IGFBP MMP-1 proHB-EGF MMP-3, -7 Cell
surface receptors FasL MMP-7 CD44 MMP-14 I
ntegrin beta3 MMP-2 Syndecan MMP-7
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Growth Growth inhibition
ECM
Latent TGFb
MMP
IGF-BP
TGFa
Survival Death
ECM
integrin
MMP
IGF-BP
HB-EGF
FasL
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Angiogenic activity Angiostatic activity
Collagen XVIII
plasminogen
Latent TGFb
Endostatin Angiostatin tumstatin
bFGF
TGFb
Collagen IV
Collagen IV
integrins
VEGF
Collagen IV Cleavage products
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Epithelial-to-mesenchymal transition
Differentiation
?
Differentiated cells
Latent TGFb
TGFb
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Invasion Invasion inhibition
Chemokine Inactivation ?
CD44
E-cadherin
Laminin-5
Collagen IV
Collagen IV Cleavage products
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MMP-TIMP balance
Development and repair Regulated, finite
imbalance
Adult, quiescence balance
Cancer dysregulated Indefinite imbalance
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cellule endothéliale
cellule tumorale
cellule immune
fibroblaste
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Tumor Metastasis